建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型

钱俞君; 秦春霞; 孙莉莉; 丁华敏; 李铁军

doi:10.3969/j.issn.1006-0111.2019.01.007

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建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型

钱俞君, 秦春霞, 孙莉莉, 丁华敏, 李铁军

文章导航 > 药学实践与服务 > 2019 > 37(1): 27-31

钱俞君, 秦春霞, 孙莉莉, 丁华敏, 李铁军. 建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型[J]. 药学实践与服务, 2019, 37(1): 27-31. doi: 10.3969/j.issn.1006-0111.2019.01.007

引用本文:

QIAN Yujun, QIN Chunxia, SUN Lili, DING Huamin, LI Tiejun. A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(1): 27-31. doi: 10.3969/j.issn.1006-0111.2019.01.007

Citation:

QIAN Yujun, QIN Chunxia, SUN Lili, DING Huamin, LI Tiejun. A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(1): 27-31. doi: 10.3969/j.issn.1006-0111.2019.01.007

建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型

doi: 10.3969/j.issn.1006-0111.2019.01.007

上海市浦东新区浦南医院药剂科, 上海 200125

基金项目: 上海市浦东新区科技发展基金（PKJ2015-Y26）；上海市浦东新区卫生和计划生育委员会学科建设（PWZbr2017-16）

A cell model for high-throughput screening lead compounds targeting HIF-1α for atherosclerosis treatment

Department of Pharmacy, Punan Hospital of Pudong New Area, Shanghai 200125, China

摘要: 目的建立一个高通量筛选防治动脉粥样硬化先导化合物的体外细胞模型。方法克隆HIF-1α低氧反应元件（hypoxia response element，HRE）至荧光素酶报告基因表达载体pGL3-Enhancer，构建荧光素酶表达载体pGL3-HIF-1α-HRE，转染人单核细胞THP-1并筛选稳定表达细胞株THP-1-HIF-1α-HRE。结果 Real Time-PCR检测表明低氧培养可以有效上调THP-1-HIF-1α-HRE细胞HIF-1α蛋白表达和荧光素酶活性，而洛伐他汀和姜黄素预处理可以有效抑制低氧引起的THP-1-HIF-1α-HRE细胞内HIF-1α蛋白表达及荧光素酶活性。结论成功建立了筛选抗动脉粥样硬化的先导化合物体外高通量细胞模型THP1-HIF-1α-HRE。
- 缺氧诱导因子1α /
- 高通量 /
- 荧光素酶活性 /
- 动脉粥样硬化 /
- 筛选
Abstract: Objective To establish a high-throughput in-vitro screening cell model for anti-atherosclerosis leading compounds. Methods Hypoxia response element (HRE) was cloned into a luciferase reporter vector, pGL3-Enhancer, to construct pGL3-HIF-1α-HRE. The THP-1 human monocyte cell line was infected with the pGL3-HIF-1α-HRE and a stable cell line, THP-1-HIF-1α-HRE, was screened. Results Real-time PCR assay showed that HIF-1α expression and luciferase activity in THP-1-HIF-1α-HRE cells was effectively upregulated by hypoxia. The increase of HIF-1α expression and luciferase activity induced by hypoxia was significantly inhibited by lovastatin or curcumin. Conclusion THP1-HIF-1α-HRE, an in-vitro cell model for high-throughput screening lead compounds for anti-atherosclerosis (AS) was successfully established.
- hypoxia inducible factor-1α /
- high-throughput /
- luciferase /
- atherosclerosis /
- screen

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建立以HIF-1α为靶标的高通量筛选防治动脉粥样硬化先导化合物的细胞模型

doi: 10.3969/j.issn.1006-0111.2019.01.007

上海市浦东新区浦南医院药剂科, 上海 200125

基金项目: 上海市浦东新区科技发展基金（PKJ2015-Y26）；上海市浦东新区卫生和计划生育委员会学科建设（PWZbr2017-16）

收稿日期: 2018-06-11
修回日期: 2018-09-25

关键词:

高通量 /

摘要: 目的建立一个高通量筛选防治动脉粥样硬化先导化合物的体外细胞模型。方法克隆HIF-1α低氧反应元件（hypoxia response element，HRE）至荧光素酶报告基因表达载体pGL3-Enhancer，构建荧光素酶表达载体pGL3-HIF-1α-HRE，转染人单核细胞THP-1并筛选稳定表达细胞株THP-1-HIF-1α-HRE。结果 Real Time-PCR检测表明低氧培养可以有效上调THP-1-HIF-1α-HRE细胞HIF-1α蛋白表达和荧光素酶活性，而洛伐他汀和姜黄素预处理可以有效抑制低氧引起的THP-1-HIF-1α-HRE细胞内HIF-1α蛋白表达及荧光素酶活性。结论成功建立了筛选抗动脉粥样硬化的先导化合物体外高通量细胞模型THP1-HIF-1α-HRE。